A stable and sensitive testing system for potential carcinogens based on DNA damage-induced gene expression in human HepG2 cell

Toxicol In Vitro. 2009 Feb;23(1):158-65. doi: 10.1016/j.tiv.2008.10.006. Epub 2008 Oct 25.

Abstract

In order to analyze potential carcinogenic and genotoxic responses caused by exposure to pollutants existing in environment, a screening method has been established in our laboratory that uses a stably transfected HepG2 cell lines containing gadd153 promoter regions which drive a luciferase reporter gene. Activation of the exogenous gadd153 promoter was quantified using the luciferase activity following drug exposure. Twenty four agents were used to evaluate this screening assay. We selected the agents, ranging from DNA alkylating agents, oxidative agent, radiation, DNA cross-linking agent, nongenotoxic carcinogens, precarcinogenic agents, which included cadmium chloride, chromium trichloride, mercuric chloride, lead nitrate, dichloro-diphenyl-trichloroethane, deltamethrin, biphenylamine, 2-aminofluorene, benzo[a]pyrene, 2,3,7,8,-tetracblorodibenzo-p-dioxin, diethyl-stilbestrol, carbon tetrachloride, mitomycin C, hydroxycamptothecin, UV, sodium fluoride, acrylamide, hydrogen peroxide. In addition, two complex genotoxic agents (water samples) existing in the environment were selected. The results showed that all 20 tested known carcinogenic and genotoxic agents were able to induce gadd153-Luc expression at a sublethal dose. In contrast, four tested non-carcinogens, included 4-acetylaminofluorene, pyrene, benzylpenicillin sodium and vitamin C, were unable to induce gadd153-Luc expression. In conclusion, this reporter system can facilitate in vitro screening for potential carcinogens. Therefore, the gadd153-Luc test system we have developed appears to be a useful and complementary system to existing genotoxic and mutagenic tests.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinogenicity Tests / methods
  • Carcinogens, Environmental / classification
  • Carcinogens, Environmental / toxicity*
  • Carcinoma, Hepatocellular / drug therapy*
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / metabolism
  • Comet Assay
  • DNA Damage
  • Dose-Response Relationship, Drug
  • Fresh Water / chemistry
  • Gene Expression / drug effects
  • Genes, Reporter / drug effects
  • Humans
  • Liver Neoplasms / drug therapy*
  • Liver Neoplasms / genetics
  • Liver Neoplasms / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Predictive Value of Tests
  • RNA, Messenger / metabolism
  • Transcription Factor CHOP / genetics
  • Transcription Factor CHOP / metabolism
  • Water Pollutants, Chemical / analysis
  • Water Pollutants, Chemical / toxicity
  • Xenobiotics / classification
  • Xenobiotics / toxicity*

Substances

  • Carcinogens, Environmental
  • DDIT3 protein, human
  • RNA, Messenger
  • Water Pollutants, Chemical
  • Xenobiotics
  • Transcription Factor CHOP
  • Luciferases